Yes / Corneal images can be acquired using confocal microscopes which provide detailed images of the different layers inside the cornea. Most corneal problems and diseases occur in one or more of the main corneal layers: the epithelium, stroma and endothelium. Consequently, for automatically extracting clinical information associated with corneal diseases, or evaluating the normal cornea, it is important also to be able to automatically recognise these layers easily. Artificial intelligence (AI) approaches can provide improved accuracy over the conventional processing techniques and save a useful amount of time over the manual analysis time required by clinical experts. Artificial neural networks (ANN) and adaptive neuro fuzzy inference systems (ANFIS), are powerful AI techniques, which have the capability to accurately classify the main layers of the cornea. The use of an ANFIS approach to analyse corneal layers is described for the first time in this paper, and statistical features have been also employed in the identification of the corneal abnormality. An ANN approach is then added to form a combined committee machine with improved performance which achieves an accuracy of 100% for some classes in the processed data sets. Three normal data sets of whole corneas, comprising a total of 356 images, and seven abnormal corneal images associated with diseases have been investigated in the proposed system. The resulting system is able to pre-process (quality enhancement, noise removal), classify (whole data sets, not just samples of the images as mentioned in the previous studies), and identify abnormalities in the analysed data sets. The system output is visually mapped and the main corneal layers are displayed. 3D volume visualisation for the processed corneal images as well as for each individual corneal cell is also achieved through this system. Corneal clinicians have verified and approved the clinical usefulness of the developed system especially in terms of underpinning the expertise of ophthalmologists and its applicability in patient care.

Yes / A confocal microscope provides a sequence of images of the various corneal layers and structures at different depths from which medical clinicians can extract clinical information on the state of health of the patient’s cornea. Preprocessing the confocal corneal images to make them suitable for analysis is very challenging due the nature of these images and the amount of the noise present in them. This paper presents an efficient preprocessing approach for confocal corneal images consisting of three main steps including enhancement, binarisation and refinement. Improved visualisation, cell counts and measurements of cell properties have been achieved through this system and an interactive graphical user interface has been developed.

No description available.

Corneal diseases and insults can result in opacification of the cornea and ultimately lead to blindness. Treatment options for patients are limited due to limited donor availability and the fact that many patients are not eligible for certain treatments due to the nature of their condition. When conventional treatment options are not beneficial for a patient, artificial corneal replacement is necessary. Current artificial replacements induce epithelial downgrowth, where the remaining host corneal cells grow underneath the replacement ultimately leading to implant extrusion. Therefore, surface modification of these synthetic materials is necessary in order to allow proper epithelialization on the surface. This work focuses on the creation of a novel corneal scaffold consisting of poly(2-hydroxyethyl methacrylate) (pHEMA) which is surface modified by 3-(acrylamido)phenylboronic acid (APBA), a molecule known to have cell-binding properties through its ability to bind sugars found throughout the cell membrane. Surfaces were modified using two different polymerization techniques: conventional free radical polymerization (CFRP) and a controlled polymerization technique known as atom transfer radical polymerization (ATRP). It was hypothesized that ATRP would yield more uniform APBA brushes than the conventional method, and therefore create a more efficient cell-binding surface than the conventional method. Following each modification, the surface chemical composition of the materials was confirmed by ATR-FTIR, XPS and surface wettability measurements. Once prepared, NIH 3T3 mouse embryo fibroblasts were seeded onto the surfaces and cell viability was assessed through an MTT assay. The results revealed no cell viability on the APBA-modified surfaces, with surface hydrophobicity, grafting density and surface toxicity (for surfaces modified through ATRP) contributing to the lack of cell attachment. / Thesis / Master of Applied Science (MASc)

Device anchoring of artificial cornea implants, through tissue integration of stromal tissue, is necessary to ensure long-term success. In this work, the delivery of basic fibroblast growth factor (FGF-2), a key modulator in corneal wound healing, via heparin modified materials was investigated as a means of sustained, soluble growth factor delivery for stimulation of device anchorage. Two materials types, commonly used for ophthalmic applications and currently under investigation for use in artificial cornea applications, were utilized. Poly (dimethyl siloxane) (PDMS) is currently under investigation as the base material for keratoprosthetic devices; dendrimer crosslinked collagen has been examined as the basis for use as a tissue engineered corneal equivalent. PDMS surfaces were modified directly or indirectly, through a poly (ethylene oxide) (PEO) spacer, to contain functionalized reactive NSC groups capable of binding heparin and FGF-2 Surface modifications were characterized with attenuated total reflection Fourier transform infrared spectrophotometer (ATR-FTIR), X-ray photoelectron spectroscopy (XPS) and water contact angles. Heparin coverage was assessed with metachromatic and bioactivity assays. Heparinized collagen gels were crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS) and polypropyleneimine octaamine G2 dendrimers. Gel integrity was assessed with water uptake, differential sr::anning calorimetry, and heparin and dendrimer stability. Both materials were exposed to radiolabelled FGF-2 and growth factor immobilization and delivery were quantified. Heparinized PDMS surfaces were capable of binding on average 100 ng/cm2 ofFGF-2, while heparinized collagen gels had higher FGF-2 immobilization, 300 ng, likely attributed to their higher heparin densities and the fact that the bulk gel rather than the surface only was modified. Delivery of FGF-2 from the heparinized materials revealed a first order release profile, with an initial burst of FGF-2, followed by gradual growth factor release. Release rates, over a 2 week period, reached 6.5% and 50%, for 1 day and 3 day FGF-2 exposed heparinized PDMS modified surfaces, while hepruinized dendrimer crosslinked collagen gels released 40%. / Thesis / Master of Applied Science (MASc)

yes / Purpose.: The aim of this work was to develop an in vitro cornea model to study the effect of proinflammatory cytokines on wound healing. Methods.: Initial studies investigated how to maintain the ex vivo models for up to 4 weeks without loss of epithelium. To study the effect of cytokines, corneas were cultured with the interleukins IL-17A, IL-22, or a combination of IL-17A and IL-22, or lipopolysaccharide (LPS). The effect of IL-17A on wound healing was then examined. Results.: With static culture conditions, organ cultures deteriorated within 2 weeks. With gentle rocking of media over the corneas and carbon dioxide perfusion, the ex vivo models survived for up to 4 weeks without loss of epithelium. The cytokine that caused the most damage to the cornea was IL-17A. Under static conditions, wound healing of the central corneal epithelium occurred within 9 days, but only a single-layered epithelium formed whether the cornea was exposed to IL-17A or not. With rocking of media gently over the corneas, a multilayered epithelium was achieved 9 days after wounding. In the presence of IL-17A, however, there was no wound healing evident. Characterization of the cells showed that wherever epithelium was present, both differentiated cells and highly proliferative cells were present. Conclusions.: We propose that introducing rocking to extend the effective working life of this model and the introduction of IL-17A to this model to induce aspects of inflammation extend its usefulness to study the effects of agents that influence corneal regeneration under normal and inflamed conditions.

yes / The cornea comprises a pool of self‐regenerating epithelial cells that are crucial to preserving clarity and visibility. Limbal epithelial stem cells (LESCs), which live in a specialized stem cell niche (SCN), are crucial for the survival of the human corneal epithelium. They live at the bottom of the limbal crypts, in a physically enclosed microenvironment with a number of neighboring niche cells. Scientists also simplified features of these diverse microenvironments for more analysis in situ by designing and recreating features of different SCNs. Recent methods for regenerating the corneal epithelium after serious trauma, including burns and allergic assaults, focus mainly on regenerating the LESCs. Mesenchymal stem cells, which can transform into self‐renewing and skeletal tissues, hold immense interest in tissue engineering and innovative medicinal exploration. This review summarizes all types of LESCs, identity and location of the human epithelial stem cells (HESCs), reconstruction of LSCN, and artificial stem cells for self‐renewal.

Corneal transplantation carries a high rate of success in the domestic cat and is an indicated treatment for specific corneal diseases in this species. The potential for iatrogenic transmission of viral diseases is a well-recognized problem in human corneal transplantation programs and screening donors for certain diseases is routine. Feline leukemia virus (FeLV) is a common agent of disease in domestic cats and available blood tests are highly effective in identification of infected individuals. This study investigates the presence of FeLV within corneal tissues of FeLV infected cats. Seventeen cats were identified to be positive for serum p27 antigen by enzyme-linked immunosorbent assay (ELISA). Twelve of these individuals were found to be positive on peripheral blood by immunofluorescent antibody (IFA) testing. Seventeen ELISA negative cats were identified to serve as negative controls. Full thickness corneal specimens were collected from all subjects and analyzed for the presence of FeLV proviral DNA and gp70 antigen by polymerase chain reaction (PCR) and immunohistochemical (IHC) testing, respectively. Eleven (64.7%) positive corneal PCR results were obtained from 17 ELISA positive cats. Of 12 cats which were both ELISA and IFA positive on peripheral blood, 10 (83.3%) had positive corneal PCR results. All corneal tissues from ELISA negative subjects were PCR negative. IHC staining of corneal sections revealed the presence of FeLV gp70 in corneal tissues of nine (52.9%) ELISA positive cats. Of the 12 cats which were both ELISA and IFA positive on peripheral blood, 8 (66.7%) had positive corneal IHC results. Positive IHC staining was localized to the corneal epithelium. Corneal tissues of all ELISA negative cats and all IFA negative cats were negative on IHC testing. This study reveals FeLV to be present within the corneal epithelium of some FeLV infected cats. Screening potential corneal donors for this virus is warranted. This work was funded by grants from the American College of Veterinary Ophthalmologists, the Virginia Veterinary Medical Association Pet Memorial Fund, and the DSACS Quick Response Fund. / Master of Science

A theoretical study of the deformation fields of the cornea under internal pressure is presented. The general elasticity equations describing a thin shell of variable thickness are solved using finite difference techniques. To gain insight into the natural corneal structure, the constant thickness case is compared to one of normal thickness. The bending stresses are found to influence the cornea's natural curvature. In the third case, the normal thickness is increased 10% to model the edematous state resulting from the incisions made during radial keratotomy. A comparison of the third case reveals the increased thickness in the peripheral cornea makes a minor contribution to the displacement; but moreover, the curvature change is opposite to that desired from radial keratotomy. The incisions are necessary to weaken the lateral support of the shell allowing the displacement and change in curvature which corrects myopia. / Master of Science

Yes / In the study of microbial keratitis, in vivo animal models often require a large number of animals, and in vitro monolayer cell culture does not maintain the three-dimensional structure of the tissues or cell-to-cell communication of in vivo models. Here, we propose reproducible ex vivo models of single- and dual-infection keratitis as an alternative to in vivo and in vitro models. / Wellcome Trust